Determination Of G-Factor - Zeiss LSM 880 Operating Manual

LSM 880 Center Screen Area / Image Containers - Display and Image Analysis
6.20.2

Determination of G-factor

In order to determine the G-factor for the LSM 710 and LSM 780 systems, you need an isotropic medium
like a fluorescent dye at a 1 mM concentration, for example fluorescein. Use the multi track configuration
with P and S analyzers selected and with a beam path and channel settings appropriate to your dye. Take
a single image and activate in the Channel tab the Ratio track. In the Ratio panel select Ratio Type 1.
Set all editable summands to 0 and all factors to 1 (see Fig. 747).
For an inverted microscope, use for convenience a glass bottom dish and pipette enough solution in to
cover the glass well. For an upright stand, squeeze a drop of solution between the glass bottom dish and
a cover slip.
If you have chosen P in the Notch filter cascade for Track 1 and assigned it to Ch1-T1 and S for track 2
assigned to Ch1-T2 then the G factor is the mean intensity value of the R1 channel. For display activate
the Histo register of the Image and select the R1 channel (see Fig. 748).
Fig. 747 Channels tool with Ratio
Type 1 formula for the
Ratio1 channel selected
The G-factor is therefore defined as:
−
Ch 1 T 1 I
= =
P
G
−
Ch 1 T 2 I
S
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CHAPTER 1 - SYSTEM OPERATION
Fig. 748
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Histogram View with Ratio1 channel
activated
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